Purification and characterization of a rabbit bone metalloproteinase that degrades proteoglycan and other connective-tissue components

Galloway, W A; Murphy, Gillian; Sandy, John D.; Gavrilović, Jelena; Cawston, Tim E.; Reynolds, John J.

doi:10.1042/bj2090741

Cited by 215 publications

(119 citation statements)

References 28 publications

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“…However, the molecular size of these enzymes on gel filtration is highly variable. In at least one case this has been attributed to the degree of purification of the enzyme [40]. Although 0.05% v/v Tween 20 was present in buffers used with the ROS 17/2 enzymes, the collagenase and gelatinase activities were not separated.…”

Section: Discussionmentioning

confidence: 99%

Synthesis of collagenase and collagenase inhibitors by osteoblast-like cells in culture

1984

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A rat osteosarcoma cell clone (ROS 17/2), and osteoblast-enriched populations from rat calvaria cultured in the presence of concanavalin A, have been shown to produce latent collagenase and collagenase inhibitors. The enzymes and inhibitor activities from the ROS 17/2 cells were concentrated by ammonium sulphate precipitation and separated by gel filtration on AcA 54 resin. The size of the latent collagenase (Mr z 58000) was reduced on conversion to active enzyme ( M , ~448000) by p-aminophenylmercuric acetate. Latent and active forms of gelatinase activity, similar in size to the corresponding forms of collagenase, were also resolved. The collagenase inhibitor activity, which was sensitive to organomercurials, was recovered in two peaks ( M , ~668000 and 30000). The active collagenase cleaved interstitial collagens (type I = I11 > 11) producing typical 3/4 and 1/4 fragments. This activity was inhibited by the metal ion chelators ethylenediaminetetraacetic acid and o-phenanthroline. Additional specific cleavages of native collagen were also observed which, from the susceptibility of this activity to phenylmethylsulphonyl fluoride, leupeptin and antipain, suggested the presence of a second collagenolytic enzyme. This synthesis of collagenolytic enzymes by these osteoblast-like cells suggests that individual osteoblasts, like fibroblasts, are capable of both synthesizing and degrading their respective organic matrices in vivo.Collagenases that characteristically cleave native interstitial collagens at neutral pH are produced by a variety of connective tissues and have been implicated in the physiological and pathological breakdown of collagen [I -31. However, whereas fibroblasts have been shown to produce collagenase in soft connective tissues, .the cellular origin of the collagenase produced by bone tissues [4-61 is equivocal. Earlier studies on bone cells derived from foetal rat calvariae have failed to demonstrate collagenase synthesis by any of several distinct populations of cells [7], which are believed to contain osteoblast-like and osteoclast-like cells [S]. However, the level of collagenase required to effect physiological remodelling of collagen could conceivably be below the detectable range of the most sensitive assays. A number of methods have been used to stimulate collagenase production by connective tissue cells to detectable levels. These include the addition of phorbol myristic acetate, cytochalasin B, bacterial endotoxins, soluble factors produced by monocytes, and collagen (reviewed by Biswas and Dayer [9]). In recent work we have shown that lectins will also stimulate collagenase production by gingival fibroblasts [lo, 111.Clonal populations of osteoblast-like cells have been isolated from foetal rat calvariae [12] and from a rat osteosarcoma [I 31. One of the osteosarcoma clones (ROS 17/2) has a stable osteoblast phenotype and has been shown to produce bone when transplanted into rats in vivo [13]. We have found that both this cell line and concanavalin-A-stimulated osteoblast-like population...

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Section: Discussionmentioning

confidence: 99%

Synthesis of collagenase and collagenase inhibitors by osteoblast-like cells in culture

1984

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“…Rheumatoid synovial fibroblasts in culture secrete three distinct matrix metalloproteinases (MMPs): MMP-1 corresponds to collagenase (EC 3.4.24.7) [1], MMP-2 to 'gelatinase' and type IV collagenase [2][3][4][5] and MMP-3 to stromelysin [6][7][8]. Collagenase digests type I, II, III and X [9] collagens.…”

Section: Introductionmentioning

confidence: 99%

“…MMP-2 is thought to be involved in the degradation of collagen by digesting gelatin derived from collagen molecules cleaved by the action of collagenase [2], although the ability of the enzyme to digest type IV and type V collagens has been pointed out [4,5]. MMP-3 has a broad range of activities to extracellular macromolecules; it degrades proteoglycans, type IV collagen, laminin, fibronectin and gelatin, and removes N-terminal propeptides of type I procollagen [6][7][8]. We have recently demonstrated that MMP-3 also digests type IX collagen which has an important role in Correspondence address: Y. Okada, Department of Pathology, School of Medicine, Kanazawa University, 13-1 Takara-machi, Kanazawa 920, Japan maintaining the structural integrity of cartilage [10].…”

Section: Introductionmentioning

confidence: 99%

Activation of matrix metalloproteinase 3 (stromelysin) and matrix metalloproteinase 2 (‘gelatinase’) by human neutrophil elastase and cathepsin G

1989

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The ability of human neutrophil elastase and cathepsin G to activate matrix metalloproteinase 3 (MMP‐3 = stromelysin) and MMP‐2 (‘gelatinase’) purified from human rheumatoid synovial fibroblasts in culture was examined. The zymogen of MMP‐3 (proMMP‐3) was activated to full activity with elastase and cathepsin G by limited proteolysis of the molecule into two active forms of M r ∼ 45000 and M r ∼ 25000. In contrast, proMMP‐2 was not activated at all by these neutrophil serine proteinases, although it was degraded into small fragments. These data suggest that neutrophil elastase and cathepsin G may play an important role in the activation of proMMP‐3 in vivo in various inflammatory conditions, but proMMP‐2 may be activated in different ways.

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“…Human leucocyte collagenase and gelatinase were prepared as published [16]. Purified rabbit bone gelatinase was a gift from C. Poll, Strangeways Laboratory (unpublished) and purified protcoglycanase was prepared by our published method [17]. Rabbit kidney metalloendopeptidase was a gift from Dr. M. Orlowski, prepared as published [18].…”

Section: Properties Of Purtiully Purified Collugenase Inhibitorsmentioning

confidence: 99%

Metalloproteinase inhibitors from bovine cartilage and body fluids

Bunning

Murphy

Kumar

et al. 1984

European Journal of Biochemistry

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Inhibitors of the mammalian metalloproteinases, collagenase, proteoglycanase and gelatinase were isolated from bovine cartilage (extracts and culture medium) and bovine amniotic fluid and serum. These inhibitors either bind or do not bind to concanavalin‐A – Sepharose, with Mr (gel filtration) of about 30000 and 20000, respectively. Cartilage and chondrocyte culture media contained only concanavalin‐A‐binding inhibitors whereas cartilage extracts contained only a non‐binding inhibitor: serum and amniotic fluid contained both forms of inhibitory activities. In most biochemical respects, particularly in their abilities to inhibit metalloproteinases, all of the inhibitors were found to be similar. It is concluded that the forms of the inhibitors that differ in Mr may be closely related to the tissue inhibitor of metalloproteinases (TIMP) previously purified from rabbit and human sources. These findings help to clarify other studies on collagenase inhibitors and support the concept that TIMP‐like inhibitors may be important in the control of connective tissue degradation.

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Purification and characterization of a rabbit bone metalloproteinase that degrades proteoglycan and other connective-tissue components

Cited by 215 publications

References 28 publications

Synthesis of collagenase and collagenase inhibitors by osteoblast-like cells in culture

Synthesis of collagenase and collagenase inhibitors by osteoblast-like cells in culture

Activation of matrix metalloproteinase 3 (stromelysin) and matrix metalloproteinase 2 (‘gelatinase’) by human neutrophil elastase and cathepsin G

Metalloproteinase inhibitors from bovine cartilage and body fluids

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